温榆河中硝化和反硝化基因的Real-timePCR定量  

Quantitative Analysis of Nitrifier and Denitrifier in Water and Sediment of the Wenyu River Using Real-time PCR

作　　者：张健伟[1,2] 孙卫玲[1,2] 邵军[1,2] 孙海美[1,2] 

机构地区：[1]北京大学深圳研究生院环境与能源学院、深圳市重金属污染控制与资源化重点实验室,广东深圳518055 [2]北京大学环境科学与工程学院,水沙科学教育部重点实验室,北京100871

出　　处：《环境科学研究》2013年第1期64-71,共8页Research of Environmental Sciences

基　　金：国家自然科学基金项目(51079002)

摘　　要：以温榆河为研究对象,采用real-time PCR研究了温榆河不同断面水样和沉积物样品中TB(总细菌)、硝化和反硝化(nosZ和narG)基因数量的变化.水样中TB基因数量在丰水期为1.05×109～7.38×1011copies/L,枯水期为1.06×109～2.69×1012copies/L;氨氧化细菌(AOB)基因数量在丰水期和枯水期分别为nd(未检出)～4.11×108和nd～1.15×109copies/L.nosZ和narG基因数量在丰水期分别为nd～2.37×108和3.61×108～1.13×1010copies/L,枯水期分别为2.0×106～3.04×109和nd～1.39×1010copies/L.枯水期沉积物样品中TB基因数量为1.35×109～7.32×1010copies/g,nosZ基因数量为nd～1.06×107copies/g,narG基因数量为1.99×107～1.02×108copies/g.枯水期TB基因数量略高于丰水期,枯水期水样中ρ(NH4+-N)较高导致其AOB基因数量要远高于丰水期,nosZ和narG基因数量并没有明显的水期变化.相关分析表明,沉积物样品中微生物基因数量与水样中微生物基因数量不相关,而是水质变化长期作用的结果.冗余度分析表明,丰水期和枯水期水样中影响微生物基因数量的主要环境因子不同,丰水期微生物基因数量是温度、ρ(CODCr)、ρ(NH4+-N)、ρ(NO2--N)、ρ(NO3--N)等共同作用的结果,而温度和ρ(CODCr)对枯水期微生物基因数量影响显著.To examine temporal and spatial changes of bacterial communities in the Wenyu River at Beijing, China, water and sediment samples were collected in wet and dry seasons from eleven sites. Quantification of total bacteria （TB）, ammonia-oxidizing bacteria （AOB） and denitrifer genes （nosZ and narG） were performed by real-time PCR. The copy numbers of the total bacteria 16S rRNA fragment were in the range of 1.05 × 109-7.38 ×lOtl copies/L in the wet season and 1.06 × 109-2.69 ×1012 copies/L in the dry season respectively. The AOB copy numbers in the wet and the dry season were nd-4. 11× 108 and nd-1. 15 × 109 copies/L respectively. The copy numbers of nosZ and narG genes were nd-2.37 × l0s and 3.61 × lO8-1. 13 × 1010 copies/L in the wet season, while they were 2.0 × 106- 3.04 × 109 and nd-1.39 ×1010 copies/L in dry season respectively. For sediments collected in the dry season, the copy numbers of TB, nosZ and narG genes were 1.35 × 109-7. 32 × 10I~ , nd-1.06 × 107 , and 1.99 × 107-1. 02 × 108 copies/g. The TB copy numbers in the dry season were slightly higher than those in the wet season, while the AOB copy numbers were much higher in the dry season than in the wet season due to high concentration of ammonia nitrogen. Pearson correlation analysis showed no significant relationshio between the bacterianumbers in the water and in the sediment. Redundancy discrimination analysis （RDA） showed that the predominant factors influencing the bacteria numbers in the wet and in the dry seasons were different. The bacteria number in the wet season was influenced by all the factors while temperature and p （ CODcr） were the main factors affecting the bacteria number in the dry

关 键 词：温榆河 氨氧化细菌 反硝化基因 real—time PCR 

分 类 号：X522[环境科学与工程—环境工程] X172